• 농업과학연구
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• 제14권2호
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• pp.286-294
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• 1987

오골계 난백 lysozyme의 용균특성을 조사하고, 이를 식품보존료로 이용하기 위한 자료를 얻고자 오골계 난백으로부터 추출 분리한 lysozyme의 각종 미생물에 대한 용균성을 조사하였다. 그 결과, 공시균중 Gram 양성균인 Staphylococcus aureus phage type 29와 Bacillus subtilis ATCC 6633에 대하여는 용균성이 인정되었으나, Gram 음성균인 E. coli를 비롯한 여타의 균종과 Staphylococcus aureus phage type 57은 lysozyme에 대한 용균 감수성이 인정되지 않았다. 분리된 lysozyme은 S. aureus phage type 29를 공시균주로 하여 이를 육즙배지에 접종하고, $37^{\circ}C$에서 24시간 정치배양하여 세포를 회수한 후 540nm에서 흡광도가 0.6이 되도록 0.05M 초산완충액(pH 4.5)에 현탁시켜, 여기에 0.05%의 lysozyme을 가하여 $30^{\circ}C$, 30분간의 조건으로 반응시킬 때 용균성이 가장 좋았다. 또한 0.05%의 lysozyme은 반응액 중에 glycine(1%)을 첨가하므로서 공시균에 대한 용균효과가 양자의 상승작용에 의하여 그 단용시보다 현저히 증대(<50%) 되었으나, 기타 다른 첨가제와 금속이온 및 lysozyme과의 혼용효과는 인정되지 않았다.

• BMB Reports
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• 제48권11호
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• pp.624-629
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• 2015

Lysozyme protects us from the ever-present danger of bacterial infection and binds to bacterial lipopolysaccharide (LPS) with high affinity. Beyond its role in the activation of protein C, the endothelial cell protein C receptor (EPCR) plays an important role in the cytoprotective pathway. EPCR can be shed from the cell surface, which is mediated by tumor necrosis factor-α converting enzyme (TACE). However, little is known about the effects of lysozyme on EPCR shedding. We investigated this issue by monitoring the effects of lysozyme on phorbol-12-myristate 13-acetate (PMA)-, tumor necrosis factor (TNF)-α-, interleukin (IL)-1βand cecal ligation and puncture (CLP)-mediated EPCR shedding and underlying mechanism. Data demonstrate that lysozyme induced potent inhibition of PMA-, TNF-α-, IL-1β-, and CLP-induced EPCR shedding. Lysozyme also inhibited the expression and activity of PMA-induced TACE in endothelial cells. These results demonstrate the potential of lysozyme as an anti-EPCR shedding reagent against PMA-mediated and CLP-mediated EPCR shedding.

• Asian-Australasian Journal of Animal Sciences
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• 제15권2호
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• pp.166-171
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• 2002

The main objective of the present study is to estimate the crossbreeding parameters in respect to serum lysozyme level in broilers. The experiment involved a complete $4{\times}4$ diallel design using four synthetic broiler lines namely Coloured Synthetic Male Line (CSML), White Synthetic Male Line (WSML), Coloured Synthetic Female Line (CSFL) and Naked Neck Line (NNL). The lyophilised Micrococcus lysodeikticus suspension was used to detect the lysozyme level in the serum of birds. The data were analysed by least-squares method to find the effects of genetic and non-genetic factors using appropriate model. The crossbreeding parameters for this trait were estimated by complete diallel model assuming the effect of each synthetic line as fixed. The results indicated that additive and non-additive genetic variation attributed to minor genes at many loci is important for the genetic control of serum lysozyme level in chickens. Total non-additive components of variance also showed significant amount of heterosis in crossbred progenies, and therefore exploitation of non-additive component of variance is possible for improvement in serum lysozyme level in broilers. The overall results suggested that for commercial broiler production system, the selection for specialised line on the basis of serum lysozyme level and subsequent crossing of parent lines could enhance the immunocompetence status in relation to serum lysozyme level in crossbred chickens.

• Animal cells and systems
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• 제12권3호
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• pp.149-155
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• 2008

We have isolated and characterized Agrius convolvuli cDNA encoding a c-type lysozyme. The cDNA sequence encodes a processed protein of 139 amino acid residues with 19 amino acid residues amino-terminal signal sequence and 120 amino acid residues mature sequence. The amino acid residues responsible for the catalytic activity and the binding of the substrate are conserved. Agrius lysozyme has a high identity to Manduca sexta. Recombinant A. convolvuli lysozyme was expressed in Escherichia coli BL21(DE3) pLysS cells for pGEX 4T-1 expression vector. Their optimal conditions for the fusion protein expression and purification were screened. Lysozyme gene amplified with primers ACLyz BamHI and ACLyz XhoI was ligated into the pGEX 4T-1 vector, which contained the glutathione S-transferase(GST) gene for fusion partner. The fusion protein was induced by IPTG and identified by SDS-PAGE analysis. Molecular weight of the fusion protein was estimated to be about 45 kDa. Recombinant lysozyme, fused to GST, was purified by glutathion-Sepharose 4B affinity chromatography. Western blot analysis of this protein revealed an immunoreactivity with the anti-Agrius lysozyme.

• 한국식품영양과학회지
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• 제16권4호
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• pp.364-370
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• 1987

한국 재래종 오골계(천연기념물 265호)의 난백으로부터 직접결정법에 의하여 lysozyme을 분리하였다. 분리된 1 % lysozyme의 Staphylococcus aureus 57, Bacillus subtilis ATCC 6633과 Escherichia coli NIHJ-JC2에 대한 항균력을 육즙 배지 속에서 lysozyme 단독으로, 또는 몇 개의 항균성 물질들과 조합하여 배양한 후 측정하였다. 그 결과, 1% lysozyme은 공시된 모든 세균에 대하여 뚜렷한 세균의 증식억제효과(약 $12{\sim}16%$를 나타내었고, 항균성 물질들과 조합하여 측정한 증식억제상의 상승효과는 S. aureus에 대해서는 0.001% magnolol과 0.001% honokiol과의 조합에서 높았고(각각 22%와 14%), E. coli에 대해서는 0.0005% erythromycin, honokiol, magnolol 그리고 phospholipase(0.005%)의 순(각각 상승효과 26,22,17,6%)으로 나타났다. 그러나, B. subtilis에 대해서는 오직 erythromycin과의 조합에서만 상승효과가 관찰되었고, 그 수준(52%)도 매우 높았다.

• Journal of Oral Medicine and Pain
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• 제39권3호
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• pp.90-95
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• 2014

Purpose: To investigate viscosity and wettability of hyaluronic acid (HA) solutions according to supplementation of lysozyme and/or peroxidase, and different ionic strength and pH conditions. Methods: Solutions containing HA were prepared using distilled deionized water (DDW) and simulated salivary buffer (SSB) in different conditions. Different concentrations of hen egg-white lysozyme and bovine lactoperoxidase was added into HA solutions. HA solutions with antimicrobials in different ionic strength and pH conditions were prepared. Viscosity was measured using cone-and-plate digital viscometer at six different shear rates and wettability on acrylic resin and Co-Cr alloy was determined by contact angle. Results: The viscosity values of HA dissolved in DDW were decreased in order of HA, HA containing lysozyme, HA containing peroxidase, and HA containing lysozyme and peroxidase. The viscosity values for HA in DDW were decreased as the concentration of lysozyme and/or peroxidase increased. However, the viscosity values for HA in SSB showed no significant changes according to the concentration of lysozyme and/or peroxidase. The viscosity values of HA solutions were inversely proportional to ionic strength and pH. The contact angle of HA solutions showed no significant differences according to tested surface materials, addition of lysozyme and/or peroxidase, and different ionic strength and pH conditions. Contact angles on acrylic resin by HA solutions in all tested conditions were much higher than those by human saliva. Conclusions: The rheological properties of HA supplemented with lysozyme and/or peroxidase in different ionic strength and pH conditions were objectively confirmed, indicating the possibility of HA with lysozyme and/or peroxidase as main components in the development of effective saliva substitutes.

• 한국식품과학회지
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• 제31권2호
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• pp.458-464
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• 1999

난백 lysozyme은 박테리아 세포벽을 선택적으로 분해하므로 식품 가공 및 저장용 천연 식품보존제로서의 이용 가치가 높다. 농도(0.25, 0.5, 1.0%, w/v)를 달리한 난백 용액을 예비여과하여 얻은 여액(PFS)과 PFS를 $35^{\circ}C$에서 한외여과하여 얻은 여액(PMS)의 단백질 농도와 lysozyme 농도를 측정하고 비활성도와 purification factor를 계산하여 최적 분리 농도를 결정하였다. 난백 용액의 각 단계별 lysozyme의 분리 정제도를 겔 크로마토그래피와 전기영동으로 확인하였으며, PM30 막에 대한 난백 단백질의 막 침착도는 SEM으로 관찰하였다. 예비여과와 한외여과 단계를 거치면서 lysozyme 이외의 비효소성 단백질은 99% 이상이 제거되었다. 비활성도는 0.25% PMS>0.50% PMS>1.0% PMS 순이었으며 PFS에 비해 PMS의 비활성도는 한외여과 단계를 거치면서 최저 18배에서 최고 31배까지 증가하였으므로, 비활성도와 purification factor가 가장 높은 0.25% 난백 용액을 최적 농도로 결정하였다. 난백 용액(0.25%, w/v)의 겔투과 크로마토그래피 결과 비효소 획분은 대부분 ovalbumin으로 판명되었으며, 전기영동 결과 PMS내에는 고순도의 lysozyme이 존재하였다. 분자량이 큰 단백질들이 $0.9{\mu}m$의 두께로 한외여과 막 표면에 침착되었으나, SEM 관찰 결과 이들 침착 단백질의 대부분은 0.1 N NaOH로 세척시 제거되는 것으로 나타났다. 따라서, polysulfone계의 PM30 막을 사용한 한외여과 공정이 고순도의 lysozyme만을 선택적으로 분리하는데 매우 효과적이었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.493-496
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• 2002

용균작용을 하는 lysozyme을 난백으로부터 분리하기 위해 이온교환크로마토그래피를 사용하였다. 용출시에 gradient를 걸어준 결과 젤과 약하게 결합된 단백질과 강하게 결합된 단백질이 분리되어 나옴을 SDS-PAGE와 Lowry methode를 통하여 알 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제9권3호
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• pp.292-296
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• 1999

The high density mixed mode adsorbent known by the trade name of Mimo-AD was used to purify lysozyme directly from the hen egg white (HEW). The homogenized hen egg white was treated with the adsorbent in a stirred vessel for lysozyme adsorption, and then the adsorbent, easily separated from the HEW by sedimentation, was packed into a column. The remaining HEW and contaminant proteins were removed by washing with pH 11 distilled water in an expanded-bed state, and subsequently the elution was performed with pH 12 distilled water in a packed-bed state. By this simple and rapid adsorption, washing, and elution procedure, lysozyme was purified to＞95％ with an overall recovery yield of 66％. This process offers a great potential for industrial application by allowing the extraction of lysozyme while retaining the commercial value of HEW.

• Preventive Nutrition and Food Science
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• 제5권2호
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• pp.65-69
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• 2000

Hen egg-white lysozyme was conjugated with 7~9 mers xyloglucan hydrolysates(MW-1,400) at 6$0^{\circ}C$ and 79% relative humidity for 3 days. SDS-PAGE showed that the conjugation between lysozyme and the oligosaccharide began from 1-day incubation, and three molecules of carbohydrate chains were attached to a protein molecule after 30day incubation. The enzymatic activity of lysozyme was totally conserved in the neoglycoprotein, when measured by using glycol chitin as substrate. Besides, the emulsifying properties of lysozyme were vastly improved by the conjugation with the oligosaccharide, in which emulsifying activity of the neoglycoprotein was five times higher than that of native one.